uwpr nano-spray-ionization sourceproteomicsresource.washington.edu/.../uwpr_nsi_source.pdf · 2015....

12/11/2013 1University of Washington Proteomics Resource

Nano-Spray-Ionization SourceAdvancing Proteomics

UWPR


UWPR NSI SourceAdvancing Proteomics

UWPR

• The NSI source can be adjusted to fit all Thermo instruments (LTQ, Orbitrap, TSQ, Fusion)

• It offers all the benefits and features of commercial NSI sources:• Lower flow rates: no drying gas or thermal heating is required and increased sensitivity compared to higher flow ESI

• Higher tolerance to a wide variety of liquid compositions than conventional ESI

• Additionally this NSI source offers significant improvements over most commercial NSI sources:• Great flexibility: it will not only accommodate in house packed columns of all length but also commercial columns, as well as

configurations with traps, spray tips etc.

• The source can be customized to your specific needs: e.g. mounting of cameras, LED lights or anything else you can think of……

• Additionally this NSI source can be used in conjunction with the Thermo NSI probes (static , dynamic and packed tip probe)

• The open design allows for leaks to be detected immediately

• Significantly lower cost

• The NSI platform can also be used with the adaptor ring of the Thermo NSI source, eliminating the need to machine the adaptorbracket

• All the parts are commercially available , except for the adaptor system, which is custom machined, e.g. by emachineshop.com

• The UWPR is currently using the NSI source on our Thermo instruments

• The plans for this NSI source are available for free, use at your own risk….

• This document and the xls file with all the part numbers should help you build your own source

• Note this is just a basic version, you can modify this source to fit your needs



UWPR

XYZ stage # 9065-XYZ

Angle plate # 9101

Rail carrier RC1

V-clamp VC1

With thumb screw # 94323A598

Fiber clamp T711-250

12” rail RLA1200

8x10” aluminum breadboard MB810

Post holder PH2

This is what we are going to build:

Construction rail CR-1

High voltage wire

Old V-clamp VC1



UWPR

XYZ stage # 9065-XYZ

Fiber clamp T711-250

12” rail # 9020

Adaptor plate

Adaptor pins

Flanged bearing

rail carrier #9025



UWPR

XYZ stage # 9065-XYZ Angle plate # 9101

Rail carrier RC1

V-clamp VC1

12” rail # 9020

rail carrier #9025

2.5” rail # 9022



UWPR

NewFocus 6” rail or 12” rail (9023 or 9020) to mount

NewFocus rail carrier (9025)

12” ThorLab rail RLA1200 to mount column using ThorLabs RC1 carrier

Note to fit the page the rail is drawn short

NewFocus Angle plate, part# 9101

ThorLab 8x10” aluminum breadboard MB810

MS

ThorLab rail carrier RC1

ThorLab V-clamp VC1

Upchurch MicroCross

NewFocus XYZ stage, part# 9065-XYZ

& 3 actuators, part# 9301 order separate

ThorLab Fiber clamp T711-250

Mounted on RC1 carrier

HV

High voltage cable; attach to cross via 0.5mm gold or platinum wire

Ion transfer tube

Adaptor pin

NewFocus rail carrier (9025)

NewPort part# CR-1

Tho

rlabs

BA

1S

Thorlabs post holder PH3

McMaster Thumb Screw 94323A598

4/27/20217University of Washington Proteomics Resource

NSI Source: step 1Advancing Proteomics

UWPR

Orbitrap FusionQuantivaTSQ AltisExploris

1. Attach base to post holder using a ¼-20 screw2. Slide the bushings into the post holder in the order shown above3. Replace the thumb screw with a ¼-20 set screw (optional)4. Lightly tighten the set screw (or thumb screw) to hold flanged bushing5. Slide the assembled post holders over the pins on the instrument

Note: you need two sets per instrument (one for each pin)

LTQLTQ-OrbitrapTSQ Vantage

2639T13

6362K233

6362K233

2706T16

2639T13

6362K233

6362K233

6362K233

6362K233

2706T16

Q Exactive seriesMSPH-2

PH2

PH3



UWPR

CR-1

5. Slide the two post holders over the two pins on the mass spectrometer6. While attached to the mass spec use two ¼-20 screws to attach the construction rail to the two bases

Make sure the construction rail is level and tighten the screws

CR-1 Construction Rail, 1 x 5 in. (Newport)

Side view

Front ViewLooking at the mass spec

CR-1

¼-20, length 1”

CR-1

PH3

PH3



UWPR

Newport 12” rail (9020NF) to mount

Newport rail carrier (9025NF) with attached 2.5” rail (9022NF)

Use two ¼”-20 x ¾” screws

to attach breadboard to CR-1

ThorLab 8x10” aluminum breadboard MB810

Use two button head screws you received with the

NewFocus rail, to attach rail to the breadboard

Use one #8-32 x ¼” with a #8-32 washer and nut and

one #8-32 x 3/8” with a nut, to attach 2.5” rail to rail carrier

Replace #8-32 setscrew

with Thumbscrew (91035A430)

CR-1

Tho

rlabs

BA

1S


9022NF e·z·Trac™ Rail, 2.5 in. Length (Newport)9025NF e·z·Trac™ Post Carrier, 0.75 in. Width (Newport)91035A430 18-8 Stainless Steel Knurled Head Captive Panel Screw (McMasterCarr)9020NF e·z·Trac™ Rail, 12.0 in. Length (Newport)MB810 Aluminum Breadboard, 8" x 10" x 1/2", ¼-20 Threaded (Thorlabs)

Newport 2.5” rail (9022NF)

Newport rail carrier (9025NF)



UWPR

Attach angle plate, part# 9101 to XYZ stage using

two ¼”-20 x 3/8” screws

Mount Newport XYZ stage, part# 9065-XYZ

To 2.5” rail using set screws provided with stage

set screws

9065-XYZ Pint-Sized XYZ-Translation Stage (Newport)AJS100-0.5K-NL High Precision Large Knob Adjustment Screw, 12.7 mm Travel, 100 TPI, No Lock (Newport)9101NF Angle Plate, 1/4-20 Thru Slots, 8-32 and 1/4-20 Threads (Newport)

Tho

rlabs

BA

1S




UWPR

Mount 12” rail RLA1200 to angle plate using two ¼”-20 x 3/8” screws

Use one #8-32 x ¼” with a #8-32 washer

To mount V clamp VC1 to ThorLab rail carrier RC1

Replace the set screw in the VC1 mounting arm with

A 8-32 nylon thumb screwUse one #8-32 x ¼” with a #8-32 washer

To mount Fiber clamp T711-250 to

ThorLab rail carrier RC1

RLA1200 Imperial Dovetail Optical Rail, 12" (Thorlabs)RC1 Rail Carrier, Counterbored Hole 1"x 1" (Thorlabs)VC1 Small V-Clamp with PM3 Clamping Arm (Thorlabs)T711-250 Post Mountable Fiber Clamp, 250mm (Thorlabs)94323A598 Nylon Raised-Head Thumb Screws (McMasterCarr)

8-32 Thread Size, 1" Long

Tho

rlabs

BA

1S



NSI Source: step 6aAdvancing Proteomics

UWPR

Jumper to bypass interlock between socket 9 and 10

DB15 male connector backside view

To bypass interlock on ThermoElectron LTQ, TSQ, QE when using alternate nanospray source (NSI)Note: Jumper and resistor can either be attached directly to the connector on the front of the LTQ or via a DB15 male connector (easier to change between different sources).

10 K ohm resistor (2% var.) for NSI source recognitionbetween socket 7 and 8

OR use DB15 male connector:solder the jumper and resistor to the corresponding pins

9 10 11 12 13 14 15

1 2 3 4 5 6 7 8

Solder Cup D-Sub Connector, DB15 Male

510-2002 Emerson Network Power, D-Sub Plug; Thermoplastic; Plug; Copper Alloy; Solder Termination (Allied Electronics)510-2445 Emerson Network Power, Hood; D-Sub; Chrome; Metalized Plastic; Steel; UL 94 VO; RoHS Compliant (Allied Electronics)70201054 Resistor; Metal Film; Res 10 Kilohms; Pwr-Rtg 0.25 W; Tol 2%; Axial; Epoxy (Allied Electronics)


NSI Source: step 6bAdvancing Proteomics

UWPR

Male Plug HARTING 09200102612: Connect 10 kΩ resistor between pin 6 and 10 and jumper wire between pin 4 and 9

Optionally a ground wire can be attached to pin 7

In case you wonder why I show it upside down… that’s because looking at the instrument from the front, that is how it will plug into the instrument

WWW

10 kΩ

X XX

X

X5 4 3 2 1

10 9 8 7 6

X

To bypass interlock on ThermoElectron Fusion or Quantiva when using alternate nanospray source (NSI)

70104585 Insert; 250 V; 16 A; Male; 10; RoHS Compliant; Han A Series; 10 A Product Size Mfr. Part#: 09200102612 (Allied Electronics)70201054 Resistor; Metal Film; Res 10 Kilohms; Pwr-Rtg 0.25 W; Tol 2%; Axial; Epoxy (Allied Electronics)



UWPR

High voltage wire:

#22 AWG, .180 O.D. silicone rubber

insulated wire (066-031-14) OR

20 AWG; 19/32; 15 kV; 0.037 in.; 0.120 in.;

200 degC; Silver Plated Copper (293-4326)

To mass spec

SCID series, threaded plastic flange receptacle

(10334-02)

Pin gold flash

over nickel (512-2474)Socket gold flash

over nickel (512-2478)

IMPORTANT: cover exposed metal with heat shrink polyolefin tubing

0.5mm gold (50822139 ) or

platinum wire (267201-400MG)

Use Upchurch micro cross or tee to connect HV to liquid path

P-777 P-875

10334-02 SCID series, threaded plastic flange receptacle assembly (Connectronics Corp)Or00004-95039 Connector, Feed thru, 15KVdc, 7.5A, Connectronics (Unity Lab Services)

066-031-14 High voltage wire, #22 AWG, .180 O.D. silicone /rubber insul. (Connectronics Corp)512-2474 20DF PIN CONT. LP (Allied Electronics)512-2478 20DF SOCKET CONT. L.P. (Allied Electronics)AA43288BU Platinum wire; 99.95% ; Alfa Aesar; 0.5mm dia; 25cm; 4.21g/m (Fisher)P-777 Microcross, .025” OD tubing sleeves, .006" THRU HOLE, PEEK (Upchurch)P-875 Microtee , w/ mount. whole, for .025” OD tubing sleeves, PEEK™ (Upchurch)F-172x Ferrule, for .025“ sleeves/P-416 NUT, PEEK™, BLACK (10 PK) (Upchurch)F-185x Tubing sleeve, 395µm (.015“) ID x .025“, PEEK™, GREEN (10 PK) (Upchurch)P-116 Plug, for micro fittings, PEEK™, BLACK (Upchurch)



UWPR

MS3R Mini Series Mounting Posts, 6mm Diameter, 3" Long (Thorlabs)MSA25 Thread Adapter, 1/4"-20 to #4-40 (Thorlabs)ER90B Mini-Post Right Angle Adapter (Thorlabs )Set screw 4-40 0.5”

Note on the Fusion and Quantiva there is a inter-lock that needs to be pushed inI used the miniature posts and a 0.5” 4-40 set screw

MS3R MSA25 ER90B

Adjust set screw to make sure the interlock is pressed in

Set screw 4-40 0.5”



UWPR

MS3R Mini Series Mounting Posts, 6mm Diameter, 3" Long (Thorlabs)MS2R Mini Series Mounting Posts, 6mm Diameter, 2" Long (ThorlabsMSA25 Thread Adapter, 1/4"-20 to #4-40 (Thorlabs)ER90B Mini-Post Right Angle Adapter (Thorlabs )4-40 screw 1in long

Note on the QE there is a inter-lock that needs to be pushed inI used the miniature posts and a 1” 4-40 set screw

MS2R MSA25 ER90B

Adjust set screw to make sure the interlock is pressed in

MS3R

Screw 4-40 1”

uwpr nano-spray-ionization sourceproteomicsresource.washington.edu/.../uwpr_nsi_source.pdf · 2015....

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